Olesen Kim, Moruzzi Noah, Bulatovic Ivana, Folmes Clifford, Jeon Ryounghoon, Felldin Ulrika, Terzic Andre, Simonson Oscar E, Le Blanc Katarina, Österholm Cecilia, Berggren Per-Olof, Schiffer Tomas, Rodin Sergey, Tilevik Andreas, Grinnemo Karl-Henrik
Department of Molecular Medicine and Surgery, Karolinska Institutet, 171 76, Stockholm, Sweden.
School of Bioscience, University of Skövde, 541 28, Skövde, Sweden.
Metabol Open. 2022 Feb 3;13:100167. doi: 10.1016/j.metop.2022.100167. eCollection 2022 Mar.
Cell metabolism has been shown to play an active role in regulation of stemness and fate decision. In order to identify favorable culture conditions for mesenchymal stromal cells (MSCs) prior to transplantation, this study aimed to characterize the metabolic function of MSCs from different developmental stages in response to different oxygen tension during expansion.
We cultured human fetal cardiac MSCs and human adult bone-marrow MSCs for a week under hypoxia (3% O) and normoxia (20% O). We performed mitochondrial characterization and assessed oxygen consumption- and extracellular acidification-rates (OCR and ECAR) in addition to oxygen-sensitive respiration and mitochondrial complex activities, using both the Seahorse and Oroboros systems.
Adult and fetal MSCs displayed similar basal respiration and mitochondrial amount, however fetal MSCs had lower spare respiratory capacity and apparent coupling efficiency. Fetal MSCs expanded in either hypoxia or normoxia demonstrated similar acidification rates, while adult MSCs downregulated their aerobic glycolysis in normoxia. Acute decrease in oxygen tension caused a higher respiratory inhibition in adult compared to fetal MSCs. In both sources of MSCs, minor changes in complex activities in normoxic and hypoxic cultures were found.
In contrast to adult MSCs, fetal MSCs displayed similar respiration and aerobic glycolysis at different O culture concentrations during expansion. Adult MSCs adjusted their respiration to glycolytic activities, depending on the culture conditions thus displaying a more mature metabolic function. These findings are relevant for establishing optimal culturing conditions, with the aim to maximize engraftment and therapeutic outcome.
细胞代谢已被证明在干细胞干性和命运决定的调节中发挥积极作用。为了确定间充质基质细胞（MSCs）移植前的适宜培养条件，本研究旨在表征不同发育阶段的MSCs在扩增过程中对不同氧张力的代谢功能。
我们在低氧（3% O₂）和常氧（20% O₂）条件下将人胎儿心脏MSCs和成人骨髓MSCs培养一周。我们使用Seahorse和Oroboros系统进行线粒体表征，并评估氧消耗率和细胞外酸化率（OCR和ECAR），以及氧敏感呼吸和线粒体复合物活性。
成人和胎儿MSCs表现出相似的基础呼吸和线粒体数量，然而胎儿MSCs的备用呼吸能力和表观偶联效率较低。在低氧或常氧条件下扩增的胎儿MSCs表现出相似的酸化率，而成人MSCs在常氧条件下下调其有氧糖酵解。与胎儿MSCs相比，氧张力的急性降低在成人MSCs中引起更高的呼吸抑制。在两种来源的MSCs中，常氧和低氧培养条件下复合物活性的变化较小。
与成人MSCs不同，胎儿MSCs在扩增过程中在不同的O₂培养浓度下表现出相似的呼吸和有氧糖酵解。成人MSCs根据培养条件将其呼吸调节为糖酵解活性，从而表现出更成熟的代谢功能。这些发现对于建立最佳培养条件具有重要意义，旨在最大限度地提高植入率和治疗效果。
